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pRK793载体质粒基本信息
pRK793质粒图谱载体图谱和pRK793载体序列质粒序列多克隆位点信息
pRK793质粒载体简介
pRK793质粒序列载体序列
LOCUS pRK793 7408 bp DNA circular SYN 12-JUL-2023
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7408)
AUTHORS Biofeng
TITLE Direct Submission
JOURNAL Exported Jul 12, 2023 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7408
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 3..80
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 81..1163
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
protein_bind 1176..1197
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1406..1434
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 1442..1458
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
CDS 1528..2628
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
regulatory 2485..2494
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2677..2688
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 2692..2712
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 2713..2733
/codon_start=1
/product="6xHis affinity tag"
/label=7xHis
/translation="HHHHHHH"
CDS 2734..3441
/codon_start=1
/product="tobacco etch virus protease (Kapust et al.,
2001)"
/label=TEV protease
/translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
EPFQPVKEATQLMN"
primer_bind complement(3498..3514)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator 3863..3949
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4041..4068
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 4088..4179
/gene="bla"
/label=AmpR promoter
CDS 4180..5040
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(5082..5595)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 5706..6294
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 6480..6620
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(6722..6913)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
1501 gcacttcacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg
1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
2461 ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg
2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaagggg agaaaatctt
2701 tattttcaag gtcatcatca tcatcatcat catggagaaa gcttgtttaa ggggccgcgt
2761 gattacaacc cgatatcgag caccatttgt catttgacga atgaatctga tgggcacaca
2821 acatcgttgt atggtattgg atttggtccc ttcatcatta caaacaagca cttgtttaga
2881 agaaataatg gaacactgtt ggtccaatca ctacatggtg tattcaaggt caagaacacc
2941 acgactttgc aacaacacct cattgatggg agggacatga taattattcg catgcctaag
3001 gatttcccac catttcctca aaagctgaaa tttagagagc cacaaaggga agagcgcata
3061 tgtcttgtga caaccaactt ccaaactaag agcatgtcta gcatggtgtc agacactagt
3121 tgcacattcc cttcatctga tggcatattc tggaagcatt ggattcaaac caaggatggg
3181 cagtgtggca gtccattagt atcaactaga gatgggttca ttgttggtat acactcagca
3241 tcgaatttca ccaacacaaa caattatttc acaagcgtgc cgaaaaactt catggaattg
3301 ttgacaaatc aggaggcgca gcagtgggtt agtggttggc gattaaatgc tgactcagta
3361 ttgtgggggg gccataaagt tttcatggtg aaacctgaag agccttttca gccagttaag
3421 gaagcgactc aactcatgaa tcgtcgtcgc cgtcgctaat aaggatcctc tagagtcgac
3481 ctgcaggcaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
3541 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
3601 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcagctt
3661 ggctgttttg gcggatgaga taagattttc agcctgatac agattaaatc agaacgcaga
3721 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
3781 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
3841 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
3901 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
3961 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
4021 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
4081 aactcttttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
4141 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
4201 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
4261 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
4321 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttc tccaatgatg
4381 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
4441 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
4501 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
4561 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
4621 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
4681 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
4741 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
4801 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
4861 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
4921 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
4981 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
5041 ctgtcagacc aagtttactc atatatactt tagattgatt taccccggtt gataatcaga
5101 aaagccccaa aaacaggaag attgtataag caaatattta aattgtaaac gttaatattt
5161 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
5221 tcggcaaaat cccttataaa tcaaaagaat agcccgagat agggttgagt gttgttccag
5281 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
5341 tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga
5401 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
5461 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
5521 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
5581 cgctacaggg cgcgtaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
5641 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
5701 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
5761 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
5821 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
5881 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
5941 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
6001 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
6061 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
6121 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
6181 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
6241 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
6301 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
6361 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
6421 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
6481 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
6541 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
6601 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
6661 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
6721 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
6781 cgtggtcgtg cagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
6841 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
6901 cctgtttggt cacttgatgc ctccgtgtaa gggggaattt ctgttcatgg gggtaatgat
6961 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
7021 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
7081 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
7141 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
7201 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
7261 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
7321 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
7381 tggccaggac ccaacgctgc ccgaaatt
//
